Discovery of lead compounds that modulate tumor specific pyruvate kinase M2 activity Abstract: We propose to discover and optimize selective modulators of human pyruvate kinase M2 (PK-M2), a splice isoform of pyruvate kinase that is specifically expressed in tumor cells. The proposed research has the following specific aims: (1) To identify both activators and inhibitors of human PK-M2 using a quantitative high-throughput screening approach against the MLSMR containing 300,000 small molecules. (2) To confirm the potency and selectivity of these compounds in a panel of secondary assays. This will include assays to identify those compounds which do not modulate the human M1, L, or R isoforms of pyruvate kinase that, in contrast to PK-M2, are expressed in most differentiated tissues but not in tumor cells. We will also determine the impact of compounds we identify on cell metabolism and proliferation. We will use these results to further improve the potency of the most promising molecules using structure-based methods, analogue synthesis and medicinal chemical principles. (3) Beyond the granting period for this proposal, to test the most promising compounds for their ability to decrease tumor growth in mouse models of cancer. PUBLIC HEALTH RELEVANCE: The M2 isoform of pyruvate kinase is expressed in all cancer cells where it plays a critical role in glucose metabolism. Because most normal tissues express another isoform of pyruvate kinase, targeting pyruvate kinase M2 provides an opportunity to selectively disrupt glucose metabolism in cancer cells. This project aims to find specific small molecule modulators of pyruvate kinase M2 to test the feasibility of targeting this enzyme in cells, and may lead to the development of novel cancer drugs. [unreadable] [unreadable] [unreadable]